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Engineering Escherichia coli for efficient and economic production of C-glycosylflavonoids by deleting YhhW and regulating pH

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Abstract

C-glycosylflavonoids have a number of pharmacological activities. An efficient method for the preparation of C-glycosylflavonoids is through metabolic engineering. Thus, it is important to prevent the degradation of C-glycosylflavonoids for producing C-glycosylflavonoids in the recombinant strain. In this study, two critical factors for the degradation of C-glycosylflavonoids were clarified. The quercetinase (YhhW) gene from Escherichia coli BL21(DE3) was expressed, purified, and characterized. YhhW effectively degraded quercetin 8-C-glucoside, orientin, and isoorientin, while the degradation of vitexin and isovitexin was not significant. Zn2+ can significantly reduce the degradation of C-glycosylflavonoids by inhibiting the activity of YhhW. pH was another key factor causing the degradation of C-glycosylflavonoids, and C-glycosylflavonoids were significantly degraded with pH exceeding 7.5 in vitro or in vivo. On this basis, two strategies, deleting YhhW gene from the genome of E. coli and regulating pH during the bioconversion, were developed to relieve the degradation of C-glycosylflavonoids. Finally, the total degradation rates for orientin and quercetin 8-C-glucoside decreased from 100 to 28% and 65% to 18%, respectively. The maximum yield of orientin reached 3353 mg/L with luteolin as substrate, and the maximum yield of quercetin 8-C-glucoside reached 2236 mg/L with quercetin as substrate. Therefore, the method described herein for relieving the degradation of C-glycosylflavonoids may be widely used for the biosynthesis of C-glycosylflavonoids in recombinant strains.

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Acknowledgements

This work was supported by the National Key R&D Program of China (2022YFD2200905) and the National Natural Science Foundation of China (21978135).

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Authors and Affiliations

  1. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China

    Simin Liu, Jiamei Liu, Linguo Zhao & Jianjun Pei

  2. Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, Nanjing, 210037, China

    Simin Liu, Jiamei Liu, Linguo Zhao & Jianjun Pei

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  1. Simin Liu
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  2. Jiamei Liu
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Correspondence to Linguo Zhao or Jianjun Pei.

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Liu, S., Liu, J., Zhao, L. et al. Engineering Escherichia coli for efficient and economic production of C-glycosylflavonoids by deleting YhhW and regulating pH. Bioprocess Biosyst Eng 46, 1251–1264 (2023). https://doi.org/10.1007/s00449-023-02893-2

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  • DOI: https://doi.org/10.1007/s00449-023-02893-2

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